LOCUS       sequence-431934-       12109 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes CAG promoter-driven dCas9-VP64 with a bipartite SV40 NLS 
            between dCas9 and VP64, along with blasticidin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12109)
  TITLE     Chavez Lab CRISPR Activator Plasmids 2024
REFERENCE   2  (bases 1 to 12109)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pb-CAGGS-dCas9-VP64-blasto
FEATURES             Location/Qualifiers
     source          1..12109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1357..1736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1738..2015
                     /label=chicken beta-actin promoter
     intron          2016..3032
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2956..2978
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3040..3059
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3129..3138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3135..3161,3162..3164,3165..5648,5649..5654,5655..5720,
                     5721..5723,5724..7238)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7251..7271
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7314..7334
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7497..7646
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     primer_bind     complement(7726..7745)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7770..7818
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        8050..8261
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8271..8280
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8277..8675
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8679..8800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8716..8735)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8770..8789
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9563..9597
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10088..10104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10088..10104)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10101..10123)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10112..10128
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(10136..10142,10143..10160,10161..10166))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10181..10202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10319..10336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10490..11078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10570..10589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11249..12040,12041..12109))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     11872..11891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccgaca ttgattattg actagttatt
     1381 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     1441 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     1501 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     1561 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     1621 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     1681 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     1741 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     1801 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc
     1861 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
     1921 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     1981 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
     2041 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     2101 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
     2161 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
     2221 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
     2281 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttttg
     2341 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     2401 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     2461 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     2521 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     2581 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     2641 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     2701 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     2761 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     2821 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     2881 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     2941 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     3001 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     3061 tgtctcatca ttttggcaaa gaatctagat accctcgtaa aggtctagag ctagcggatc
     3121 gaacccttgc caccatggac aagaagtact ccattgggct cgctatcggc acaaacagcg
     3181 tcggctgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc aaagttctgg
     3241 gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg ttcgactccg
     3301 gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat acccgcagaa
     3361 agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag gtggatgact
     3421 ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag cacgagcgcc
     3481 acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac ccaaccatat
     3541 atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg ttgatctatc
     3601 tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg gacctgaacc
     3661 cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac aatcagcttt
     3721 tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg agcgctaggc
     3781 tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag aagaagaacg
     3841 gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt aaatctaact
     3901 tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat gatgatctcg
     3961 acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg gcaaagaacc
     4021 tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc accaaagctc
     4081 cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg actttgctga
     4141 aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc gatcagtcta
     4201 aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt tacaaattta
     4261 ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag cttaacagag
     4321 aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac cagattcacc
     4381 tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt ttgaaagata
     4441 acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta ggccccctcg
     4501 cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc atcactccct
     4561 ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc gaaaggatga
     4621 ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct ctgctgtacg
     4681 agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa gggatgagaa
     4741 agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc ttcaagacga
     4801 accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt gaatgtttcg
     4861 actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga acgtatcacg
     4921 atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac gaggacattc
     4981 ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt gaagaacgct
     5041 tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag aggcgccgat
     5101 atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac aagcagagtg
     5161 gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac ttcatgcagt
     5221 tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa gtttctggcc
     5281 agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct atcaaaaagg
     5341 gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga aggcataagc
     5401 ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag ggacagaaga
     5461 acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg tcccaaatcc
     5521 ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac ctgtactacc
     5581 tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg ctctccgact
     5641 acgacgtggc tgctatcgtg ccccagtctt ttctcaaaga tgattctatt gataataaag
     5701 tgttgacaag atccgataaa gctagaggga agagtgataa cgtcccctca gaagaagttg
     5761 tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc acacaacgga
     5821 agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat aaagccggct
     5881 tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc caaattctcg
     5941 attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag gtgaaagtta
     6001 ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt tataaggtga
     6061 gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg gtaggcactg
     6121 cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac tataaagtgt
     6181 acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc accgctaagt
     6241 acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg gccaatggag
     6301 agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc gtgtgggaca
     6361 agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg aacatcgtta
     6421 aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca
     6481 gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc ggattcgatt
     6541 ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg aagtctaaaa
     6601 aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca agcttcgaaa
     6661 aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca
     6721 ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta
     6781 gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac gttaatttct
     6841 tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat gagcagaagc
     6901 agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa ataagcgaat
     6961 tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct gcttacaata
     7021 agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg tttactctga
     7081 ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac agaaagcggt
     7141 acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt acggggctct
     7201 atgaaacaag aatcgacctc tctcagctcg gtggagacag cagggctgac cccaagaaga
     7261 agaggaaggt gggttctgga aaacggacag cggacggtag cgagtttgag agtccgaaga
     7321 aaaagaggaa agtagagggt ggttctgccg gtggctccgg ttctggctcc agcggtggca
     7381 gctctggtgc gtccggcacg ggtactgcgg gtggcactgg cagcggttcc ggtactggct
     7441 ctggcgctgt acggtcgggt ggctctggat cagaggccag cggttccgga cgggctgacg
     7501 cattggacga ttttgatctg gatatgctgg gaagtgacgc cctcgatgat tttgaccttg
     7561 acatgcttgg ttcggatgcc cttgatgact ttgacctcga catgctcggc agtgacgccc
     7621 ttgatgattt cgacctggac atgctgatta actctagagg tgtacagtga taacctccta
     7681 gggcggaaga acactgctag gtgagtttaa acgggggagg ctaactgaaa cacggaagga
     7741 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     7801 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     7861 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     7921 ccccaagttc gggtgaaggc ccagggctcg cagccaatgt cggggcggca ggccctgcca
     7981 tagcagatct gcgaaatcgg atccgcggcc gcactgattc tagctgcaga ggctccggtg
     8041 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     8101 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     8161 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     8221 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggatcctcga gccaccatgg
     8281 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
     8341 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
     8401 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
     8461 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
     8521 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
     8581 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
     8641 aattgctgcc ctctggttat gtgtgggagg gctaagttaa cttgtttatt gcagcttata
     8701 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     8761 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg aattgactca
     8821 aatgatgtca attagtctat cagaagctca tctggtctcc cttccggggg acaagacatc
     8881 cctgtttaat atttaaacag cagtgttccc aaactgggtt cttatatccc ttgctctggt
     8941 caaccaggtt gcagggtttc ctgtcctcac aggaacgaag tccctaaaga aacagtggca
     9001 gccaggttta gccccggaat tgactggatt ccttttttag ggcccattgg tatggctttt
     9061 tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa
     9121 gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat
     9181 gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg
     9241 ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctga tatctataac
     9301 aagaaaatat atatataata agttatcacg taagtagaac atgaaataac aatataatta
     9361 tcgtatgagt taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg
     9421 cggtcgttat agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag
     9481 ctccaagcgg cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag
     9541 agagagcaat atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta
     9601 atctagctgc atcaggatca tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc
     9661 tggcgctatc tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg
     9721 gcatggaaag agtttgccga ggatgactgc tgctgcattg acgttgagcg aaaacgcacg
     9781 tttaccatga tgattcggga aggtgtggcc atgcacgcct ttaacggtga actgttcgtt
     9841 caggccacct gggataccag ttcgtcgcgg cttttccgga cacagttccg gatggtcagc
     9901 ccgaagcgca tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg
     9961 cagattaatg acagcggtgc ggcgctggga tattacgtca gcgaggacgg gtatcctggc
    10021 tggatgccgc agaaatggac atggataccc cgtgagttac ccggcgggcg cgcttggcgt
    10081 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
    10141 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
    10201 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
    10261 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
    10321 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
    10381 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
    10441 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
    10501 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
    10561 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
    10621 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
    10681 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
    10741 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
    10801 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
    10861 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
    10921 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
    10981 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
    11041 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
    11101 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
    11161 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
    11221 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
    11281 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
    11341 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
    11401 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
    11461 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
    11521 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
    11581 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
    11641 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
    11701 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
    11761 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
    11821 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
    11881 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
    11941 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
    12001 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
    12061 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcat
//
